Bernard Lam's Biography

Bernard Lam,
Senior Research Scientist,
Norgen Biotek Corporation

Next Generation Sequencing Characterization of Small RNA “Signatures” of Different Bodily Fluids and the Practical Implications for Biomarker Discovery Studies

Wednesday, 28 March 2018 at 16:30

Add to Calendar ▼2018-03-28 16:30:002018-03-28 17:30:00Europe/LondonNext Generation Sequencing Characterization of Small RNA “Signatures” of Different Bodily Fluids and the Practical Implications for Biomarker Discovery StudiesSELECTBIOenquiries@selectbiosciences.com

In this presentation, we will discuss the impact of various steps in the
Next Generation Sequencing workflow on obtaining good small RNA
expression profile from a specific bodily fluid. Topics discussed will
include RNA extraction, enhancement of target signal by abundant
transcript removal and library preparation. The use of bodily fluids
such as whole blood, plasma, serum, urine or saliva for biomarker
discovery has attracted tremendous attention in recent years. However,
the characteristics of RNA (and DNA) present in bodily fluids are quite
different from traditional samples such as cells and tissues. Many of
the RNAs in bodily fluids are either free-circulating or are within
extra-cellular vesicles such as exosomes. More importantly, the RNAs
present in bodily fluids are usually of very low abundance and small in
size. This presents challenges for detection and discovery using many
next-generation gene expression technologies. Using deep sequencing, we
estimated the maximum number of each small RNA species in different
bodily fluids. A guideline to how much read depth is required for each
sample type will be discussed. We will present data on the “signatures”
or small RNA expression characteristics of different bodily fluids. We
will present examples, using these RNA expression “signatures” of a
bodily fluid, to show that removing an abundant RNA transcript can
significantly improve the small RNA diversity detected by sequencing.
Finally, we will discuss different RNA extraction technologies and their
ability to recover microRNA with low GC contents from bodily fluids
with low RNA content.